Fresh stacks of muddy bacteria

Lilia Garcia, Illinois Wesleyan University

The Approach: In my last post, I wrote about Gracilaria, an invasive red seaweed on the coast of South Carolina, and its effect on Vibrio bacteria. My project aims to record the number and strains, or types, of Vibrio growing around Gracilaria and compare it to seaweed-free areas. I will also compare the Vibrio count residing on Gracilaria versus the Vibrio residing on a native seaweed called Ulva to see how an invasive species changes the bacterial community. Lastly, I want to understand how Gracilaria stops the growth of specific Vibrio strains by producing chemical compounds.

Mud samples under Gracilaria, taken by K. Coates

To begin solving my questions, I will go out to collect samples in the mudflats outside of the Grice Laboratory. I will collect tubes of water, clumps of Gracilaria and Ulva, and mud from underneath and 1.5 feet away from Gracilaria. Afterwards, I’ll spread all the samples onto dishes with nutrients specifically used to grow Vibrio. The bacteria grow in spots called colonies, and I will count each spot to see how much Vibrio there is in each sample. I am looking for a different amount of colonies in mud samples collected within or away from Gracilaria patches, and a difference in colony numbers between the Ulva and Gracilaria.

Dishes of unique Vibrio, taken by L. Garcia

A single dish from a mud sample can contain hundreds of colonies, differing in color, shape, size, and texture. Each of these colonies represent a different strain of Vibrio, uncovering the diversity of bacteria at different distances from Gracilaria. I will characterize which unique colonies are dangerous to human health, and whether they are found near or away from Gracilaria.

Zones of inhibition against Vibrio strain, taken by L. Garcia

As previously mentioned, I will also test Vibrio strains against chemical compounds made on the surface of Gracilaria. These compounds are able to control the kind of bacteria that grow around seaweed, changing the microscopic habitat. I will mix Gracilaria with chemicals to remove its surface chemistry, then spot the compounds onto dishes growing Vibrio from my mud samples. I am looking for large clear circles, called zone of inhibitions, that tell me the specific strain of Vibrio cannot grow due to the compound.

Nearly all we know about the ecological and economic impact of Gracilaria focuses on large animals, such as fish. My project zooms in on micro-organisms that have been overlooked. The information I collect will help us understand how invasive Gracilaria is changing bacterial communities not only in the Charleston Harbor, but potentially the entire coast.  Although invisible, bacteria make up the foundation of ecosystems and high Vibrio levels may be dangerous for our health. I look forward to finding the answers to my questions hiding quietly in the mud.

Acknowledgements

Thank you to my mentor Dr. Erik Sotka, and our collaborator Dr. Erin Lipp. I would also like to thank Dr. Alan Strand and Kristy Hill-Spanik for their supporting guidance. Lastly, thank you to Dr. Loralyn Cozy (IWU) for preparing me to succeed in the lab. All research is funded by Grice Marine Lab and College of Charleston through the Fort Johnson REU Program, NSF DBI-1757899

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Invisible Neighbors: How Gracilaria Changes Bacterial Communities

Lilia Garcia, Illinois Wesleyan University

The Problem: It only takes a walk along the mudflats to notice large patches of wiry, red seaweed. The seaweed is called Gracilaria vermiculophylla, an invasive organisms that is native to East Asia (SERC, 2019)  The seaweed is hard to miss, but its effects on the ecosystem are not easily seen. This summer I will be studying how Gracilaria affects a bacterial community invisible to the naked eye.

Mudflat with Gracilaria, taken by L. Garcia

According to previous studies, Gracilaria is found to increase the amount of a bacteria called Vibrio (Gonzalez, et al., 2014). This may not mean much at first, since most of us don’t think about microscopic interactions. Bacteria, however, are important in maintaining the health of complex environments like estuaries. They cycle and break down nutrients and organic matter, influencing oxygen, carbon, and nitrogen levels. An increase in one group of bacteria, such as Vibrio, can change these patterns. And like most of us know, bacteria tends to spread easily. There are a few strains, or types, of Vibrio, such as V. vulnificus, V. parahaemolyticus, and V. cholera, that are dangerous to human health. An increase in these strains may cause an increase in disease from swimming or eating infected food.†

Vibrio growing on petri dish, taken by L. Garcia

We known Vibrio levels increase with Gracilaria, but we do not know how this happens. We also don’t know if all Vibrio strains increase together, or if only a few strains grow. To understanding the relationship between Gracilaria and Vibrio, I will record how much total Vibrio and how many strains of Vibrio grow in and away from patches of Gracilaria. In order to preserve its own health, Gracilaria produces compounds that promote or stop organisms from growing around it (Assaw et al., 2018). These are compounds I will test against different strains to study the mechanism Gracilaria uses affect specific Vibrio levels. I want to see how the growth of each strain is affected by different extracts. Will the strains further away from the Gracilaria be unable to grow when exposed to a certain type of extract? Will other strains grow better with the extract?

We tend to think about invasive species on a large scale, assessing the damage it causes to other familiar animals and plants. The ecosystem relies on tiny, cellular organism and studying how bacteria changes leads to a deeper understanding of environmental health. An invisible community is changing as Gracilaria flourishes, and there is a lot left to learn about it. 

Acknowledgements

Thank you to my mentor Dr. Erik Sotka, and our collaborator Dr. Erin Lipp. I would also like to thank Dr. Alan Strand and Kristy Hill-Spanik for their supporting guidance. Lastly, thank you to Dr. Loralyn Cozy (IWU) for preparing me to succeed in the lab. All research is funded by Grice Marine Lab and College of Charleston through the Fort Johnson REU Program, NSF DBI-1757899

References

Assaw S, Rosli N, Adilah N, Azmi M, Mazlan N, Ismail N. 2018. Antioxidant and Antibacterial Activities of Polysaccharides and Methanolic Crude Extracts of Local Edible Red Seaweed Gracilaria sp. Malays Appl Biol. 47(4): 135-144. 

Fofonoff PW, Ruiz GM, Steves B, Simkanin C, & Carlton JT. 2019. National Exotic Marine and Estuarine Species Information System. 

Gonzalez D, Gonzalez R, Froelich B, Oliver J, Noble R, McGlathery K. 2014. Non-native macroalga may increase concentrations of Vibrio bacteria on intertidal mudflats. Mar Ecol Prog Ser. 505: 29-36.

Catch of the Day(s)

Melanie Herrera, University of Maryland College Park

South Carolina is known for its iconic southern cuisine, including a staple of fresh seafood which fuels the buckets of shrimp & grits and “catch of the day”. In order to support this huge industry (and fill the bellies of every South Carolinian), I am conducting an experiment to figure out where this seafood is holing up prior to its demise. Dr. Harold, his graduate student, Mary Ann McBrayer, and I are out on Grice Beach collecting fishes, crabs, shrimps, and much more in order to figure what exactly is there… And what they are using to survive.

Using a seine net, we encircle marine animals in dense and sparse patches of an invasive sea grass, Gracilaria, for collection. We hypothesize that Gracilaria is helping the local economy (a surprising contribution from an invasive species) by creating refuge for young animals. On the beach, we submerge separate samples of animals (from dense versus sparse areas of Gracilaria) into a euthanizing solution to bring them up to the lab for preservation and analysis (Figure 1).

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Figure 1: An example of animals caught in separate habitats at Grice Cove. The left exhibits animals caught in a dense area of Gracilaria and the right exhibits animals caught from a sparse area of Gracialaria. Credit: Melanie Herrera

In the lab, separate samples (dense versus sparse) undergo a few transfers into different fixatives (10% seawater formalin, 25% isopropyl, and 50% isopropyl consecutively) to keep the fish from decaying. After this preserving process, fish and other animals are separated and categorized by family, genus, and species. This categorization enables us to identify and analyze what types of animals and how many of each are using different habitat. Our analysis will give us insight on what type of habitat, either patches dominated by Gracilaria or areas with more open water, benefits fish. Specifically, we will be able to identify if Gracilaria is more advantageous to young fish or if their survivorship is independent from their habitat.

So far, we have collected lots of pipefish, narrow skinny fish that resemble a hair strand-size snake, Atlantic Silversides, a fish that looks exactly like it sounds, and more shrimp than anyone needs (Figure 2). Although some of these animals do not directly contribute to the seafood industry, its presence in the Charleston Harbor can tell us a lot of things. For example, we have seen some fishes that usually stay in warmer waters in the Southern U.S. Their expanding habitat can lead us to some more hypotheses on climate change and warm weather moving northward. In addition, we can find out if Gracilaria has a stake in rearing economically important fish in the future.

Pipefish, Silversides, Grass Shrimp.png

Figure 2: (From left to right) Pipefish, Atlantic Silversides, and Grass Shrimp caught for analysis.Credit: Melanie Herrera

Thank you so much to my mentor Dr. Tony Harold and his lab for his advice and guidance. Thank you to Mary Ann McBrayer for helping me facilitate this project. This research is funded through the National Science Foundation and College of Charleston’s Grice Marine Lab.

 

Invasive Species: Friend or Foe

Melanie Herrera,  University of Maryland – College Park

Invasive species…. Haunting, domineering, and downright evil. Or are they? Unlike the infamous Zebra Mussels, dominating the Great Lakes, or Fire Ants, constantly wreaking havoc, Gracilaria Vermiculophylla, are giving invasive species a good name. Don’t get me wrong, invasive species infuriate me just as much as the next guy; but Dr. Tony Harold and I are here to draw out the benefits of this invasive sea grass to baby fish.

Unlike the native, simpler sea grass previously occupying Charleston Harbor, Gracilaria is characterized by coarse branching structures that appeal to many species of fish as protective homes. We are particularly interested in fishes in the larval and juvenile stages (the young ones) that associate with these complex habitats. Having access to more protective sea grass, such as this invasive, in these vulnerable life stages can help determine how many of these little guys make it into adulthood. Similar macro-algae to Gracilaria, such as seaweeds, have been known to be preferable hideouts for larvae and juveniles, reducing the pressures of predation. Since Gracilaria is on the rise in our local estuary, the Charleston Harbor, it’s important to find out the role they play in keeping our fish alive and well.

Our project is designed to better understand the level of association of local fish such as Gobies, Atlantic Menhaden, Atlantic Silversides, and other estuary-occupying fishes, with Gracilaria. We will compare abundance and distribution of young fish in dense patches of Gracilaria to sparse patches. Maybe these young fishes prefer the familiarity that native sea grass and open water brings. Or maybe Gracilaria’s “new and improved” design is too advantageous to resist. After we figure this out, we can go on sustainably managing local fish critical to commercial and recreational use and condemning the rest of the invasive species.

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An example of a collection site characterized as a “dense” habitat of Graclaria vermiculophylla.  Photo Credit: Melanie Herrera

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An example of a collection site characterized as a “sparse” habitat of Gracilaria vermiculophylla. Photo Credit: Melanie Herrera

 

Thank you so much to my mentor Dr. Tony Harold and his lab for his advice and guidance. Thank you to Mary Ann McBrayer for helping me facilitate this project. This research is funded through the National Science Foundation and College of Charleston’s Grice Marine Lab.

 

Works Cited

Munari, N. Bocchi & M. Mistri (2015) Epifauna associated to the introduced Gracilaria vermiculophylla (Rhodophyta; Florideophyceae: Gracilariales) and comparison with the native Ulva rigida (Chlorophyta; Ulvophyceae: Ulvales) in an Adriatic lagoon, Italian Journal of Zoology, 82:3, 436-445, DOI: 10.1080/11250003.2015.1020349